title: Neuropeptides to replace serum in cryopreservation of mesenchymal stromal cells?
abstract: OBJECTIVE
  The therapeutic potential of human mesenchymal stromal cells (MSCs) has generated considerable interest in a wide variety of areas. MSC banking is feasible, but the optimal technique of cryopreservation remains to be determined.
  
  
  METHODS
  To reduce dimethyl sulfoxide (DMSO) concentration in cryopreservation medium, DMSO was replaced with sucrose or trehalose. To increase cell survival and proliferation rates after thawing and to eliminate the need for fetal bovine serum (FBS), neuropeptides of the vasoactive intestinal peptide/glucose-dependent insulinotropic peptide/pituitary adenylate cyclase activating polypeptide family were added to the cryopreservation medium. Cell survival was analyzed by a trypan blue dye exclusion assay. Cell proliferation of cryopreserved MSCs was determined after 7 days of culture.
  
  
  RESULTS
  No significant differences in cell survival rates were detected between cryopreservation solutions with 5% and 10% DMSO, independently of the addition of trehalose or sucrose. Cell proliferation rates tended to be highest when MSCs were frozen in 5% DMSO + trehalose. FBS could be replaced by human albumin (HA) without loss in cell survival and proliferation potential. With FBS, the addition of neuropeptides could increase cell survival and proliferation rates. Without FBS or HA, cell survival and proliferation rates in the presence of neuropeptides were comparable to rates achieved with FBS or HA.
  
  
  CONCLUSIONS
  Classic cryopreservation with 10% DMSO could be replaced by 5% DMSO + 30 mmol/L trehalose. FBS could be replaced by HA or neuropeptides without loss in cell survival and proliferation potential. The addition of neuropeptides in the cryopreservation medium containing FBS could increase the cell proliferation rate and consequently cellular output.
authors.affiliation: Giga-I(3) Research, Hematology Unit, CHU of Liege and University of Liege, Liege, Belgium.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Briquet
authors.forename: Amélie
authors.initials: A
authors.lastname: Halleux
authors.forename: Chantal
authors.initials: C
authors.lastname: Lechanteur
authors.forename: Yves
authors.initials: Y
authors.lastname: Beguin
mesh_terms: D018070:Biological Specimen Banks; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide; D005615:Freezing; D005749:Gastric Inhibitory Polypeptide / Q000494:pharmacology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide / Q000494:pharmacology*; D014660:Vasoactive Intestinal Peptide / Q000494:pharmacology*
chemical_list: D003451:Cryoprotective Agents; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D014660:Vasoactive Intestinal Peptide; D005749:Gastric Inhibitory Polypeptide; D004121:Dimethyl Sulfoxide
keywords: GIP; PACAP; VIP; cryopreservation; mesenchymal stromal cells; neuropeptides